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Papers In Press, published online ahead of print February 19, 2002
J. Biol. Chem, 10.1074/jbc.M107962200
Submitted on August 17, 2001
Revised on February 19, 2002
Accepted on February 19, 2002

Human prepro-orexin gene regulatory region that activates gene expression in the lateral region and represses it in the medial region of the hypothalamus

Takashi Moriguchi, Takeshi Sakurai, Satoru Takahashi, Katsutoshi Goto, and Masayuki Yamamoto

Molecular and Developmental Biology, Center for Tsukuba Advanced Research Alliance, Tsukuba City 305-8577

Corresponding Author: masi{at}tara.tsukuba.ac.jp

Prepro-orexin is a precursor of the neuropeptides orexin-A and -B that are localized in the neuronal population of the lateral hypothalamic area (LHA). We wish to elucidate the mechanisms by which the prepro-orexin gene is specifically activated in orexin neurons of the LHA. The 3.2-kb 5’-flanking region of the human prepro-orexin gene is sufficient for the specific expression of an E. coli LacZ reporter gene in orexin neurons. We, therefore, examined a series of reporter constructs harboring this 3.2-kb regulatory region or its deletion in a reporter transgenic mouse assay. There are two phylogenetically conserved regions located 287-bp (OE1) and 2.5-kb (OE2) upstream of the transcription initiation site of the human prepro-orexin gene. In transgenic mice, both OE1 and OE2 are necessary for expressing the human prepro-orexin gene in the LHA and repressing its expression in the medial regions of the hypothalamus. Through a serial deletion analysis of OE1, we found that the 57-bp core region of OE1 is critical for its spatial gene regulatory function in vivo. A mutation analysis further demonstrated that without contribution from the OE1 core region, the LacZ reporter is expressed ectopically in the medial regions of the hypothalamus. Thus, OE1 contains crucial cis-acting elements regulating prepro-orexin gene expression specifically in the LHA.


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